Improved production and in vivo assembly of soluble...

C - Chemistry – Metallurgy – 12 – N

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C12N 7/04 (2006.01) C07K 14/08 (2006.01) C12P 21/02 (2006.01)

Patent

CA 2685308

The present invention provides an improved method for the in vivo production of soluble assembled virus-like particles ('VLPs') in bacterial cells of Pseudomonad origin. The Pseudomonad cells support assembly of VLPs from icosahedral viral capsid proteins ('CPs') in vivo, and allow the inclusion of larger recombinant peptides as monomers or concatamers in the VLP. The invention specifically provides an improved method for the in vivo production of soluble assembled Cowpea Chlorotic Mottle Virus ('CCMV') VLPs by introducing modifications into the CCMV CP that result in high yield production of soluble CP fusions in a Pseudomonas fluorescens bacterial system. These soluble VLPs can subsequently be purified and used as vaccines.

L'invention concerne un procédé amélioré pour la production in vivo de particules assemblées solubles analogues à un virus ('VLP') dans des cellules bactériennes issues de Pseudomonades. Les cellules de Pseudomonades permettent d'assembler des VLP provenant de protéines capsidiques virales icosaédriques ('CP') in vivo, et d'inclure des peptides recombinés plus grands comme monomères ou concatamères dans le VLP. L'invention concerne spécifiquement un procédé amélioré pour la production in vivo de VLP solubles assemblées du virus 'CCMV', par l'introduction, dans la CP du CCMV, de modifications engendrant une production à haut rendement de fusions des CP solubles dans un système bactérien fluorescent de Pseudomonas. Ces VLP solubles peuvent être ultérieurement purifiées et utilisées comme vaccins.

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